title: The controlled generation of functional basal forebrain cholinergic neurons from human embryonic stem cells.
abstract: An early substantial loss of basal forebrain cholinergic neurons (BFCN) is a constant feature of Alzheimer's disease and is associated with deficits in spatial learning and memory. The ability to selectively control the differentiation of human embryonic stem cells (hESCs) into BFCN would be a significant step toward a cell replacement therapy. We demonstrate here a method for the derivation of a predominantly pure population of BFCN from hESC cells using diffusible ligands present in the forebrain at developmentally relevant time periods. Overexpression of two relevant human transcription factors in hESC-derived neural progenitors also generates BFCN. These neurons express only those markers characteristic of BFCN, generate action potentials, and form functional cholinergic synapses in murine hippocampal slice cultures. siRNA-mediated knockdown of the transcription factors blocks BFCN generation by the diffusible ligands, clearly demonstrating the factors both necessary and sufficient for the controlled derivation of this neuronal population. The ability to selectively control the differentiation of hESCs into BFCN is a significant step both for understanding mechanisms regulating BFCN lineage commitment and for the development of both cell transplant-mediated therapeutic interventions for Alzheimer's disease and high-throughput screening for agents that promote BFCN survival.
authors.affiliation: Department of Neurology, Northwestern University's Feinberg School of Medicine, Chicago, Illinois, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Bissonnette
authors.forename: Ljuba
authors.initials: L
authors.lastname: Lyass
authors.forename: Bula J
authors.initials: BJ
authors.lastname: Bhattacharyya
authors.forename: Abdelhak
authors.initials: A
authors.lastname: Belmadani
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Miller
authors.forename: John A
authors.initials: JA
authors.lastname: Kessler
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D004594:Electrophysiology; D053595:Embryonic Stem Cells / Q000166:cytology*; D055427:Growth Differentiation Factor 2 / Q000494:pharmacology; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D060850:LIM-Homeodomain Proteins; D051379:Mice; D018613:Microscopy, Confocal; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D016548:Prosencephalon / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D014157:Transcription Factors; D014212:Tretinoin / Q000494:pharmacology
chemical_list: C422443:GBX1 protein, human; D055427:Growth Differentiation Factor 2; D018398:Homeodomain Proteins; C112960:LIM homeobox protein 8; D060850:LIM-Homeodomain Proteins; D014157:Transcription Factors; D014212:Tretinoin